Docetaxel albumin nanoparticle pharmaceutical composition, preparation method therefor and use thereof

ABSTRACT

The present invention relates to a docetaxel albumin nanoparticle pharmaceutical composition, a preparation method therefor, and a use thereof for manufacturing drugs for treating cancer. The pharmaceutical composition comprises docetaxel, albumin and amino acid(s), wherein the weight ratio between albumin and docetaxel is no more than 50, preferably is 20:1 to 1:1, and the weight ratio between amino acid(s) and docetaxel is not less than 0.5, preferably is 1:1 to 20:1.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a national stage filing under 35 U.S.C. § 371of PCT/CN2016/077521, filed on 28 Mar. 2016, and claims priority toChinese patent application 201510157393.1, filed 3 Apr. 2015. Eachapplication is incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

The present invention pertains to the field of pharmaceuticalcompositions of docetaxel (docetaxol), specifically relates to adocetaxel albumin nanoparticle pharmaceutical composition, preparationmethod therefor and use thereof, and especially relates to a docetaxelalbumin nanoparticle pharmaceutical composition with improved stability,preparation method therefor and use thereof.

BACKGROUND OF THE INVENTION

Docetaxel belongs to taxane drugs. The main mechanism of action thereofinvolves interfering with mitosis, promoting the assembly ofmicrotubules and prohibiting the disassembly of microtubules, so as toinhibit the differentiation of tumor cells, and finally lead to deaththereof. Currently, docetaxel is approved for the treatment of variousindications such as breast cancer, non-small cell lung cancer, prostatecancer and the like in major countries worldwide, and it constitutes themost commonly used or standard therapy for the treatment of thesecancers. Furthermore, in subsequent clinical studies, docetaxel is alsowidely investigated for the treatment of gastric cancer, head and necktumor, esophageal cancer, ovarian cancer, and the like. Treatment ofthese indications with docetaxel is expected to be approved in Europeand America later.

Docetaxel has poor water solubility. Presently, the formulation ofdocetaxel commercially available is a liquid dosage form for infusion ina concentrated form, and the dosage regimen normally recommended isintravenous infusion of 75 mg/m² within 1 hour, once every 3 weeks. Adocetaxel injection comprises a drug concentrate, which is formed bydissolving docetaxel in a solvent, Tween-80, accompanied with a diluentcontaining 13% ethanol. However, pre-application of an anti-allergicdrug is necessary during clinical administration due to the hemolyticproperty of Tween-80, which tends to cause allergic reactions includingdyspnea, hypotension, angioedema, rubella, shock, etc. when it isintravenously injected. Moreover, the high viscosity of Tween-80 alsobrings about great inconvenience in clinical application.

In order to overcome defects of the docetaxel injection, such as toxicside effects and the like, CN103830181A discloses a lyophilized liposomecomprising an inclusion complex of docetaxel with cyclodextrin, whereinthe stability of docetaxel is improved by inclusion with cyclodextrin,and an improved targeting property and reduced toxic side effects areachieved with the liposome particle system. However, the toxicity ofcyclodextrin per se limits the application thereof. CN101773465Adiscloses a polymeric micellar system stabilized by amino acids, and apolymeric micelle comprising docetaxel is developed. It is shown thatthe physical appearance of the polymeric micelle with amino acids can bestable for more than 5 days, while the polymeric micelle without aminoacids can be stable only for 30 minutes. However, the degradation ofhigh molecular polymers (e.g., mPEG-PLA, etc.) employed in polymericmicelles after being injected into a body is quite slow, and may evenlast for more than 1 year. In view of such potential safety issues, nopolymeric micelle product of docetaxel has been approved for marketingby the FDA in the United States. As such, although improved targetingproperties and reduced toxic side effects are achieved with theseparticle systems, the application thereof is limited by their defects.

CN103054798A discloses a docetaxel albumin nanoparticle (ABI-008),wherein citric acid (or a salt thereof) is added to a composition ofdocetaxel and albumin, such that the physical stability of a solution ofthe docetaxel albumin nanoparticle is increased, and no precipitation orsedimentation phenomenon is observed for at least 8 hours afterreconstitution or rehydration.

However, for improving the stability of docetaxel, in addition tocontrol the physical stability of particles in a solution, it is moreimportant to reduce the chemical degradation of docetaxel. At present,studies focusing on reducing the chemical degradation of docetaxel arequite limited in the prior art, and no method for improving suchdegradation is available yet.

Docetaxel per se can undergo a variety of degradation pathways undervarious conditions, and degradation products resulted therefrom rendercorresponding changes in the activity and/or toxicity of docetaxel, andmay even significantly affect the activity and/or toxicity thereof. Themain factors affecting the degradation of docetaxel include temperature,acidic and basic solvents, oxidants, reductants and light, etc.

In a basic, neutral or strong acidic medium, one of the main degradationpathways of docetaxel is the epimerization of 7-hydroxy, which resultsin 7-epi-docetaxel through retro-aldol reaction.

Bornique et al. (Drug Metabolism and Disposition, Vol. 30, No. 11, pp.1149-1152, 2002) investigates the interaction between docetaxel and7-epi-docetaxel with recombinant human cytochrome P4501B1 (hCYP1B1).hCYP1B1 is a common cytochrome in human tumor cells, and is mainlyrelated to drug resistance of chemotherapy drugs (including docetaxel).The in vitro test shows that the activity of hCYP1B1 can be increased bymore than 7 times by 7-epi-docetaxel, thus it is confirmed that thedegradation product of docetaxel, 7-epi-docetaxel, reduces the activityof docetaxel.

CN101415416A discloses inhibition of the production of 7-epi-docetaxelin a pharmaceutical composition of docetaxel and polysorbate 80 byadding an organic acid with a pKa value of 2.5 to 4.5 as a docetaxeldegradation inhibitor.

However, the inventors of the present invention have demonstratedthrough experimentation that addition of an agent such as tartaric acid,citric acid, ascorbic acid or another organic acid with a pKa value of2.5 to 4.5 etc. into a composition of docetaxel and albumin cannoteffectively inhibit the production of 7-epi-docetaxel, which, instead,may even increase. This indicates that the above agents decrease thechemical stability of the composition, and affect the safety of thefinal formulation.

CN103054798A discloses addition of a stabilizer such as citric acid (ora salt thereof) and the like into a composition of docetaxel andalbumin. Although the stabilizer enhances the physical stability of thesolution of the docetaxel albumin nanoparticle, the inhibitory effectthereof on the production of 7-epi-docetaxel is not mentioned.

It has been demonstrated through experimentation that conventionalstabilizers such as citric acid (or a salt thereof) and the like cannotalways effectively inhibit the production of 7-epi-docetaxel, andsometimes the production thereof may be accelerated. During storage, theamount of 7-epi-docetaxel produced far exceeds 2.0%, which would resultsin potential safety issues in clinical medication.

One of the challenges in search for a method for inhibiting theproduction of 7-epi-docetaxel is that, during the storage of a docetaxelalbumin nanoparticle, the presence of a large amount of hydrogen bondsin the polypeptidic structure of albumin is unfavorable for thestability of docetaxel and accelerates the retro-aldol reaction, whichresult in the epimerization of 7-hydroxyl, rendering the formation andsteady increase of the 7-epi-docetaxel impurity.

As such, it is an urgent problem to be solved in the art to find amethod for inhibiting the production of 7-epi-docetaxel in apharmaceutical composition of docetaxel.

SUMMARY OF THE INVENTION

In view of the defects in the prior art, according to an aspect of thepresent invention, a docetaxel albumin nanoparticle pharmaceuticalcomposition with enhanced stability is provided, wherein thepharmaceutical composition comprises:

docetaxel,

albumin, and

amino acid(s),

wherein the weight ratio of albumin to docetaxel is not greater than 50,i.e., is from 50:1 to >0:1, preferably is from 50:1 to 1:1, and morepreferably is from 20:1 to 1:1.

According to another aspect of the present invention, the abovedocetaxel albumin nanoparticle pharmaceutical composition is provided,wherein the weight ratio of the amino acid(s) to docetaxel is no lessthan 0.5, i.e., is ≥0.5:1, preferably is from 0.5:1 to 80:1, morepreferably is no less than 1, i.e., is ≥1:1, and preferably is from 1:1to 20:1.

According to another aspect of the present invention, the abovedocetaxel albumin nanoparticle pharmaceutical composition is provided,wherein the pharmaceutical composition further comprises a proteinstructure unfolding agent.

According to another aspect of the present invention, a method forpreparing the above docetaxel albumin nanoparticle pharmaceuticalcomposition is provided.

According to another aspect of the present invention, a formulationcomprising the above docetaxel albumin nanoparticle pharmaceuticalcomposition is provided, and the formulation further comprises apharmaceutically acceptable carrier and/or auxiliary material.

According to another aspect of the present invention, a use of the abovepharmaceutical composition or the above formulation in the manufactureof a medicament for the treatment of cancer is provided.

According to another aspect of the present invention, a method fortreating cancer is provided, wherein the method comprises administeringto a subject in need thereof an effective amount of the abovepharmaceutical composition or the above formulation.

According to another aspect of the present invention, the abovepharmaceutical composition or the above formulation for the treatment ofcancer is provided.

Surprisingly, it is found that the addition of amino acid(s) can resultin a docetaxel albumin nanoparticle pharmaceutical composition which isstable for a long time, and can significantly inhibit the production of7-epi-docetaxel in the pharmaceutical composition. According to anembodiment of the present invention, addition of amino acid(s) into adocetaxel albumin nanoparticle pharmaceutical composition can result ina composition with a content of 7-epi-docetaxel below 1% after beingstored for 24 months (e.g. at 2-8° C.). Particularly, when the aminoacid employed is arginine, the content of 7-epi-docetaxel is preferablybelow 1%, such as about 0.98%, about 0.83%, about 0.75%, about 0.72% oreven lower, after 30 months of storage (e.g. at 2-8° C.).

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 shows the pharmacological efficacy results of the composition ofExample 1-c in the treatment of a subcutaneous xenograft of humannon-small cell lung cancer cell A549 in a tumor model of BALB/c nudemice.

DETAILED DESCRIPTION OF THE INVENTION

Pharmaceutical Composition and Preparation Method Therefor

An embodiment of the present invention provides a docetaxel albuminnanoparticle pharmaceutical composition, wherein the pharmaceuticalcomposition comprises:

docetaxel,

albumin, and

amino acid(s),

wherein the weight ratio of albumin to docetaxel is not greater than 50,i.e., is from 50:1 to >0:1, preferably is from 50:1 to 1:1, and morepreferably is from 20:1 to 1:1.

A further embodiment of the present invention provides the abovedocetaxel albumin nanoparticle pharmaceutical composition, wherein theweight ratio of the amino acid(s) to docetaxel is no less than 0.5,i.e., is ≥0.5:1, preferably is from 0.5:1 to 80:1, more preferably is noless than 1, i.e., is ≥1:1, and preferably is from 1:1 to 20:1.

A further embodiment of the present invention provides the abovedocetaxel albumin nanoparticle pharmaceutical composition, which furthercomprises a protein structure unfolding agent including, e.g., one ormore of mercaptoethanol, glutathione, acetylcysteine, anddithiothreitol, wherein the weight ratio of albumin to the proteinstructure unfolding agent is not greater than 100:1, i.e., is from 100:1to >0:1, such as from 90:1 to >0:1, from 80:1 to >0:1, from 70:1to >0:1, from 60:1 to >0:1, from 50:1 to >0:1, from 40:1 to >0:1, from30:1 to >0:1, from 20:1 to >0:1 or from 10:1 to >0:1, and preferably isfrom 50:1 to >0:1.

A further embodiment of the present invention provides the abovedocetaxel albumin nanoparticle pharmaceutical composition, wherein thepharmaceutical composition is a nanoparticle suspension, which comprisesdocetaxel in a concentration of at least 1 mg/ml, and the nanoparticlestherein have a particle diameter of not greater than 200 nm.

Another embodiment of the present invention provides a method forpreparing the above docetaxel albumin nanoparticle pharmaceuticalcomposition, wherein the method comprises the following steps:

1) dissolving docetaxel in an organic solvent preferably selected fromone or more of ethanol, tert-butyl alcohol, and acetone, to obtain anorganic phase;

2) using an aqueous solution containing amino acid(s) to dissolvealbumin or to dilute a solution of albumin, so as to obtain a solutionof albumin and amino acid(s);

3) adding a protein structure unfolding agent to the solution of albuminand amino acid(s) obtained in step 2), and performing an incubationreaction; wherein the protein structure unfolding agent includes one ormore of mercaptoethanol, glutathione, acetylcysteine, anddithiothreitol;

4) adding the organic phase obtained in step 1) to the solution obtainedafter the incubation reaction in step 3) under shear, to obtain a dilutesolution of docetaxel albumin nanoparticles; and

5) concentrating the solution obtained in step 4) by ultrafiltration, toobtain a docetaxel albumin nanoparticle pharmaceutical composition withenhanced stability;

wherein the weight ratio of albumin to docetaxel is not greater than 50,i.e., is from 50:1 to >0:1, preferably is from 50:1 to 1:1, and morepreferably is from 20:1 to 1:1; the weight ratio of the amino acid(s) todocetaxel is no less than 0.5, i.e., is ≥0.5:1, preferably is from 0.5:1to 80:1, more preferably is no less than 1, i.e., is ≥1:1, andpreferably is from 1:1 to 20:1; and the weight ratio of albumin to theprotein structure unfolding agent is not greater than 100:1, i.e., isfrom 100:1 to >0:1, and preferably is from 50:1 to >0:1.

Another embodiment of the present invention provides a method forpreparing the above docetaxel albumin nanoparticle pharmaceuticalcomposition, wherein the method comprises the following steps:

1) dissolving docetaxel in an organic solvent preferably selected fromone or more of ethanol, tert-butyl alcohol, and acetone, to obtain anorganic phase;

2) using an aqueous solution containing amino acid(s) to dissolvealbumin or to dilute a solution of albumin, so as to obtain a solutionof albumin and amino acid(s);

3) subjecting the solution of albumin and amino acid(s) obtained in step2) to an incubation reaction;

4) adding the organic phase obtained in step 1) to the solution obtainedafter the incubation reaction in step 3) under shear, to obtain a dilutesolution of docetaxel albumin nanoparticles; and

5) concentrating the solution obtained in step 4) by ultrafiltration, toobtain a docetaxel albumin nanoparticle pharmaceutical composition withenhanced stability;

wherein the weight ratio of albumin to docetaxel is not greater than 50,i.e., is from 50:1 to >0:1, preferably is from 50:1 to 1:1, and morepreferably is from 20:1 to 1:1; and the weight ratio of the aminoacid(s) to docetaxel is no less than 0.5, i.e., is ≥0.5:1, preferably isfrom 0.5:1 to 80:1, more preferably is no less than 1, i.e., is ≥1:1,and preferably is from 1:1 to 20:1.

Another embodiment of the present invention provides a method forpreparing the above docetaxel albumin nanoparticle pharmaceuticalcomposition, wherein the method comprises the following steps:

1) dissolving docetaxel in an organic solvent preferably selected fromone or more of ethanol, tert-butyl alcohol, and acetone, to obtain anorganic phase;

2) using water for injection to dissolve albumin or to dilute a solutionof albumin, so as to obtain a solution of albumin;

3) adding a protein structure unfolding agent to the solution of albuminobtained in step 2), and performing an incubation reaction; wherein theprotein structure unfolding agent includes one or more ofmercaptoethanol, glutathione, acetylcysteine, and dithiothreitol;

4) adding the organic phase obtained in step 1) to the solution obtainedafter the incubation reaction in step 3) under shear, to obtain a dilutesolution of docetaxel albumin nanoparticles; and

5) concentrating the solution obtained in step 4) by ultrafiltration,adding amino acid(s) to the concentrate, wherein the weight ratio of theamino acid(s) to docetaxel is no less than 0.5, i.e., is ≥0.5:1,preferably is from 0.5:1 to 80:1, more preferably is no less than 1,i.e., is ≥1:1, and preferably is from 1:1 to 20:1, to obtain a docetaxelalbumin nanoparticle pharmaceutical composition with enhanced stability;

wherein the weight ratio of albumin to docetaxel is not greater than 50,i.e., is from 50:1 to >0:1, preferably is from 50:1 to 1:1, and morepreferably is from 20:1 to 1:1; and the weight ratio of albumin to theprotein structure unfolding agent is not greater than 100:1, i.e., isfrom 100:1 to >0:1, and preferably is from 50:1 to >0:1.

Another embodiment of the present invention provides a method forpreparing the above docetaxel albumin nanoparticle pharmaceuticalcomposition, wherein the method comprises the following steps:

1) dissolving docetaxel in an organic solvent preferably selected fromone or more of ethanol, tert-butyl alcohol, and acetone, to obtain anorganic phase;

2) using water for injection to dissolve albumin or to dilute a solutionof albumin, so as to obtain a solution of albumin;

3) subjecting the solution of albumin obtained in step 2) to anincubation reaction;

4) adding the organic phase obtained in step 1) to the solution obtainedafter the incubation reaction in step 3) under shear, to obtain a dilutesolution of docetaxel albumin nanoparticles; and

5) concentrating the solution obtained in step 4) by ultrafiltration,adding amino acid(s) to the concentrate, wherein the weight ratio of theamino acid(s) to docetaxel is no less than 0.5, i.e., is ≥0.5:1,preferably is from 0.5:1 to 80:1, more preferably is no less than 1,i.e., is ≥1:1, and preferably is from 1:1 to 20:1, to obtain a docetaxelalbumin nanoparticle pharmaceutical composition with enhanced stability;

wherein the weight ratio of albumin to docetaxel is not greater than 50,i.e., is from 50:1 to >0:1, preferably is from 50:1 to 1:1, and morepreferably is from 20:1 to 1:1.

Another embodiment of the present invention provides a method forpreparing the above docetaxel albumin nanoparticle pharmaceuticalcomposition, wherein the method comprises the following steps:

1) dissolving docetaxel in an organic solvent preferably selected fromone or more of ethanol, tert-butyl alcohol, and acetone, to obtain anorganic phase;

2) using water for injection to dissolve albumin or to dilute a solutionof albumin, so as to obtain a solution of albumin;

3) adding amino acid(s) and a protein structure unfolding agent to thesolution of albumin obtained in step 2), and performing an incubationreaction; wherein the protein structure unfolding agent includes one ormore of mercaptoethanol, glutathione, acetylcysteine, anddithiothreitol;

4) adding the organic phase obtained in step 1) to the solution obtainedafter the incubation reaction in step 3) under shear, to obtain a dilutesolution of docetaxel albumin nanoparticles; and

5) concentrating the solution obtained in step 4) by ultrafiltration, toobtain a docetaxel albumin nanoparticle pharmaceutical composition withenhanced stability;

wherein the weight ratio of albumin to docetaxel is not greater than 50,i.e., is from 50:1 to >0:1, preferably is from 50:1 to 1:1, and morepreferably is from 20:1 to 1:1; the weight ratio of the amino acid(s) todocetaxel is no less than 0.5, i.e., is ≥0.5:1, preferably is from 0.5:1to 80:1, more preferably is no less than 1, i.e., is ≥1:1, andpreferably is from 1:1 to 20:1; and the weight ratio of albumin to theprotein structure unfolding agent is not greater than 100:1, i.e., isfrom 100:1 to >0:1, and preferably is from 50:1 to >0:1.

Another embodiment of the present invention provides a method forpreparing the above docetaxel albumin nanoparticle pharmaceuticalcomposition, wherein the method comprises the following steps:

1) dissolving docetaxel in an organic solvent preferably selected fromone or more of ethanol, tert-butyl alcohol, and acetone, to obtain anorganic phase;

2) using water for injection to dissolve albumin or to dilute a solutionof albumin, so as to obtain a solution of albumin, and then adding aminoacid(s);

3) subjecting the solution of albumin obtained in step 2) to anincubation reaction;

4) adding the organic phase obtained in step 1) to the solution obtainedafter the incubation reaction in step 3) under shear, to obtain a dilutesolution of docetaxel albumin nanoparticles; and

5) concentrating the solution obtained in step 4) by ultrafiltration, toobtain a docetaxel albumin nanoparticle pharmaceutical composition withenhanced stability;

wherein the weight ratio of albumin to docetaxel is not greater than 50,i.e., is from 50:1 to >0:1, preferably is from 50:1 to 1:1, and morepreferably is from 20:1 to 1:1; and the weight ratio of the aminoacid(s) to docetaxel is no less than 0.5, i.e., is ≥0.5:1, preferably isfrom 0.5:1 to 80:1, more preferably is no less than 1, i.e., is ≥1:1,and preferably is from 1:1 to 20:1.

According to the above embodiments, amino acid(s) can be added before orafter the formation of nanoparticles during the preparation of thedocetaxel albumin nanoparticle pharmaceutical composition of the presentinvention. During the preparation of the present composition, any methodcan achieve the effect of the present invention, as long as it can mixdocetaxel albumin nanoparticles with amino acid(s). The above-listedmethods are merely major ones, and without departing from the spirit ofthe present invention, other methods obtained through reasonablemodifications to the above methods are covered by the present invention.

Unless otherwise defined in the context, all technical and scientificterms used herein are intended to have the same meaning as commonlyunderstood by a person skilled in the art. References to techniquesemployed herein are intended to refer to the techniques as commonlyunderstood in the art, including variations on those techniques orsubstitutions of equivalent techniques which would be apparent to aperson skilled in the art. While it is believed that most of thefollowing terms will be readily understood by a person skilled in theart, the following definitions are put forth to better illustrate thepresent invention.

The terms “contain”, “include”, “comprise”, “have”, or “involve”, aswell as other variations thereof used herein are inclusive oropen-ended, and do not exclude additional, unrecited elements or methodsteps.

As used herein, the term “docetaxel” is also referred to as “Taxotere”or “docetaxel” in the art, and the “docetaxel” referred to in thepresent application comprises the docetaxel compound per se andderivatives or analogues thereof. The derivatives or analogues ofdocetaxel include but are not limited to compounds similar to docetaxelin structure, or compounds which belong to a same generic chemicalspecies as that of docetaxel, such as taxanes. In some embodiments, thederivatives or analogues of docetaxel possess biological,pharmacological, chemical and/or physical properties (including, e.g.,functionality) similar to those of docetaxel. Examples of thederivatives or analogues of docetaxel include paclitaxel andcabazitaxel. Moreover, as used herein, the term “docetaxel” alsoincludes crystalline and amorphous forms, as well as anhydrite, hydratedforms (such as hemihydrate, dihydrate, trihydrate, and the like) andsolvated forms (such as alcoholate) thereof.

As used herein, the term “albumin” includes one or more of recombinantalbumin and serum albumin, and the serum albumin includes non-humananimal (e.g., bovine) serum albumin and human serum albumin, preferablyhuman serum albumin.

As used herein, the term “amino acid(s)” includes at least one of basicpolar amino acids (such as arginine or lysine), nonpolar amino acids(such as proline), neutral polar amino acids (e.g., cysteine) and acidicpolar amino acids (e.g., aspartic acid or glutamic acid), preferablyarginine.

As used herein, the term “protein structure unfolding agent” refers to asubstance that can unfold hydrophobic bond regions of albumin tofacilitate the binding of docetaxel with albumin. All the substanceswith such an effect can achieve the object of the present invention. Theprotein structure unfolding agent includes but is not limited to one ormore of mercaptoethanol, glutathione, acetylcysteine and dithiothreitol.

In a preferred embodiment, the pharmaceutical composition is ananoparticle suspension, which comprises docetaxel in a concentration ofat least 1 mg/ml, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 25, 30, 35,40 or 50 mg/ml, and the nanoparticles therein have a particle diameterof not greater than 200 nm, e.g., 150, 100, 95, 90, 85, 80, 75, 70, 65,60, 55 or 50 nm. The particle diameter of the nanoparticles and theconcentration of docetaxel can be in any range comprised of the abovevalues of the particle diameter or concentration.

The pharmaceutical composition of the present invention retains desiredtherapeutic effects, and remains physically and/or chemically stablewhen exposed to certain conditions, such as prolonged storage, elevatedtemperature or dilution for parenteral administration.

The term “physically stable” means that no substantial precipitation orsedimentation phenomenon is observed for at least about 8 hours,preferably within about 24 hours, more preferably within 48 hours, andparticularly preferably within 96 hours, after reconstitution orrehydration of a lyophilized powder formulation of the pharmaceuticalcomposition of the present invention.

The term “chemically stable” means that the chemical structure orcomposition of the pharmaceutical composition or active compound isstable when the pharmaceutical composition of the present invention isstored under conventional conditions. Preferably, after storage at 2-8°C. for at least 24 months, or even up to 30 months, the contentpercentage of 7-epi-docetaxel is ≤1.0%.

Pharmaceutical Formulation and Therapeutic Method

Another embodiment of the present invention provides a formulationcomprising a therapeutically effective amount of the above docetaxelalbumin nanoparticle pharmaceutical composition and a pharmaceuticallyacceptable carrier and/or auxiliary material. The formulation ispreferably a lyophilized powder formulation.

Another embodiment of the present invention provides a use of thepharmaceutical composition or formulation of the present invention inthe manufacture of a medicament for the treatment or prevention of anabnormal cell proliferative disease or disorder. The abnormal cellproliferative disease or disorder is preferably cancer, which includesprostate cancer, colon cancer, breast cancer, head and neck cancer,pancreatic cancer, lung cancer and ovarian cancer.

Another embodiment of the present invention provides a use of thepharmaceutical composition or formulation of the present invention inthe manufacture of a medicament for the treatment or prevention ofcancer, including prostate cancer, colon cancer, breast cancer, head andneck cancer, pancreatic cancer, lung cancer and ovarian cancer.

Another embodiment of the present invention provides a method fortreating or preventing an abnormal cell proliferative disease ordisorder, wherein the method comprises administering to a subject inneed thereof a therapeutically effective amount of the pharmaceuticalcomposition or formulation of the present invention. The abnormal cellproliferative disease or disorder is preferably cancer, and the canceris preferably a specific cancer listed above.

The term “pharmaceutically acceptable carrier” refers to a diluent,auxiliary material, excipient, or vehicle with which a therapeutic isadministered, and it is, within the scope of sound medical judgment,suitable for contact with the tissues of human beings and animalswithout excessive toxicity, irritation, allergic response, or otherproblem or complication, commensurate with a reasonable benefit/riskratio.

The pharmaceutically acceptable carrier which can be employed in theformulation of the present invention includes, but is not limited tosterile liquids, such as water and oils, including those of petroleum,animal, vegetable or synthetic origin, such as peanut oil, soybean oil,mineral oil, sesame oil and the like. Water is an exemplary carrier whenthe formulation is administered intravenously. Physiological salines aswell as aqueous dextrose and glycerol solutions can also be employed asliquid carriers, particularly for injectable solutions. Suitablepharmaceutical excipients include starch, glucose, lactose, sucrose,gelatin, maltose, chalk, silica gel, sodium stearate, glycerolmonostearate, talc, sodium chloride, dried skim milk, glycerol,propylene glycol, water, ethanol and the like. The formulation, ifdesired, can also contain minor amounts of wetting or emulsifyingagents, or pH buffering agents. Oral formulations can include standardcarriers such as pharmaceutical grades of mannitol, lactose, starch,magnesium stearate, sodium saccharine, cellulose, magnesium carbonate,etc. Examples of suitable pharmaceutical carriers are described in e.g.Remington's Pharmaceutical Sciences (1990).

The formulation can act systemically and/or locally. To this end, it canbe administered through a suitable route, such as through injection,intravenous, intraarterial, subcutaneous, intraperitoneal,intramuscular, or transdermal administration, or administered via oral,buccal, nasal, transmucosal, topical, as an ophthalmic formulation, orvia inhalation.

For these routes of administration, the formulation can be administeredin a suitable dosage form.

Such dosage form includes, but is not limited to a spray, lotion,ointment, suspension, injectable solution, suspension injection,emulsion injection, elixir, syrup or lyophilized powder formulation.

As used herein, the term “therapeutically effective amount” refers tothe pharmaceutical composition/formulation being administered which willrelieve to some extent one or more of the symptoms of the disorder beingtreated.

Dosage regimens may be adjusted to provide the optimum desired response.For example, a single bolus may be adminstered, several divided dosesmay be administered over time or the dose may be proportionally reducedor increased as indicated by the exigencies of the therapeuticsituation. It is to be noted that dosage values may vary with the typeand severity of the condition to be alleviated, and may include singleor multiple doses. It is to be further understood that for anyparticular subject, specific dosage regimens should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of thecompositions.

The amount of the pharmaceutical composition/formulation of the presentinvention administered will be dependent on the subject being treated,the severity of the disorder or condition, the rate of administration,the disposition of the pharmaceutical composition/formulation and thediscretion of the prescribing physician. Generally, an effective dosageis in the range of about 0.0001 to about 50 mg per kg body weight perday, for example about 0.01 to about 10 mg/kg/day, in single or divideddoses. For a 70 kg human, this would amount to about 0.007 mg to about3500 mg/day, for example about 0.7 mg to about 700 mg/day. In someinstances, dosage levels below the lower limit of the aforesaid rangemay be more than adequate, while in other cases still larger doses maybe employed without causing any harmful side effect, provided that suchlarger doses are first divided into several small doses foradministration throughout the day.

The term “treating”, as used herein, unless otherwise indicated, meansreversing, alleviating, inhibiting the progress of, or preventing thedisorder or condition to which such term applies, or one or moresymptoms of such disorder or condition.

As used herein, the term “subject” includes a human or non-human animal.An exemplary human subject includes a human subject having a disease(such as one described herein) (referred to as a patient), or a normalsubject. The term “non-human animal” as used herein includes allvertebrates, such as non-mammals (e.g. birds, amphibians, reptiles) andmammals, such as non-human primates, livestock and/or domesticatedanimals (such as sheep, dog, cat, cow, pig and the like).

Advantageous Effects of the Pharmaceutical Composition of the PresentInvention

1. The amino acid(s) added significantly inhibits the formation of the7-epi-docetaxel impurity in a docetaxel albumin nanoparticlepharmaceutical composition, enhances the stability, especially chemicalstability, of the docetaxel nanoparticle pharmaceutical composition,prolongs the shelf life of the docetaxel nanoparticle pharmaceuticalcomposition, reduces side effects or toxicity upon administration,decreases irritation to an organism, and improves clinical tolerability.

2. The method for preparing the pharmaceutical composition of thepresent invention is simple.

3. The present invention can be employed for the treatment of variouscancers, such as prostate cancer, colon cancer, breast cancer, head andneck cancer, pancreatic cancer, lung cancer (especially non-small celllung cancer) and ovarian cancer, etc., and thus has a broad prospect ofapplication on the market. Moreover, the docetaxel albumin nanoparticlepharmaceutical composition of the present invention has a bettertherapeutic effect than that of a commercially available docetaxelinjection.

EXAMPLES

The following examples are provided to further illustrate the presentinvention. It is necessary to be pointed out that the following examplesshould not be construed as limiting the protection scope of the presentinvention, and technical solutions obtained through certainnon-substantial improvements and modifications to the technicalsolutions of the present invention by a skilled artisan according to thecontents described above are still covered by the present invention.

The concentration of the commercially available docetaxel injectionemployed in the examples of the present application was 40 mg/ml. Theterm “docetaxel” in the examples is the docetaxel (i.e., docetaxel)compound per se.

Example 1

An organic phase was obtained by adding 100 ml of ethanol into 1500 mgof docetaxel, which was then dissolved by sonication. Human serumalbumin was diluted with water for injection to formulate a 6 mg/mlsolution of human serum albumin. 500 mg of glutathione was added to 1250ml of the 6 mg/ml solution of human serum albumin, and the solution wasincubated in a water bath at about 70° C. for 6 minutes to obtain anaqueous phase. The organic phase was homogeneously dispersed into theaqueous phase at high-speed shear (1,000 revolutions per minute (rpm)),and the resulting system was transferred to an equipment forconcentration by ultrafiltration (PALL, 100 kD membrane). The system wasconcentrated by ultrafiltration to obtain a 6 mg/ml concentrate ofdocetaxel, wherein the docetaxel albumin nanoparticles had an averageparticle diameter of 103 nm (Malvern Nano-ZS90).

The above docetaxel albumin nanoparticle concentrate was divided intothree portions, and arginine was added in the following ratios:

1-a. the weight ratio of arginine to docetaxel was 0.5:1;

1-b. the weight ratio of arginine to docetaxel was 1:1; and

1-c. the weight ratio of arginine to docetaxel was 20:1.

The mixed solutions containing arginine were then respectivelysterilized by filtration through a 0.22 μm sterile filter head, andfreeze-dried for 48 hours in a lyophilizer.

The lyophilized samples were reconstituted with physiological saline,and no precipitation was observed when the reconstituted samples wereleft at room temperature for 8 hours, indicating that the docetaxelalbumin nanoparticle pharmaceutical composition of the present inventionwas physically stable. The particle diameters were measured (the averageparticle diameter of sample 1-a was 103 nm, the average particlediameter of sample 1-b was 102 nm, and the average particle diameter ofsample 1-c was 104 nm), and the increase trends of the 7-epi-docetaxelimpurity in the docetaxel albumin nanoparticle pharmaceuticalcompositions were determined.

Example 2

An organic phase was obtained by adding 10 ml of ethanol into 150 mg ofdocetaxel, which was then dissolved by sonication. Bovine serum albuminsolids were dissolved with water for injection to formulate a 6 mg/mlsolution of bovine serum albumin. 200 mg of glutathione and 3750 mg ofproline were added to 1250 ml of the 6 mg/ml solution of bovine serumalbumin, and the solution was incubated in a water bath at about 60° C.for 6 minutes to obtain an aqueous phase. The organic phase washomogeneously dispersed into the aqueous phase at high-speed shear(1,000 revolutions per minute (rpm)), and the resulting system wastransferred to an equipment for concentration by ultrafiltration (PALL,100 kD membrane). The system was concentrated by ultrafiltration toobtain a 6 mg/ml concentrate of docetaxel, wherein the finalconcentration of proline was 3 mg/ml, and the weight ratio of proline todocetaxel was about 0.5:1.

The docetaxel albumin nanoparticles had an average particle diameter of113 nm (Malvern Nano-ZS90). The concentrate obtained was sterilized byfiltration through a 0.22 μm sterile filter head, and freeze-dried for48 hours in a lyophilizer. The lyophilized sample was reconstituted withphysiological saline, and no precipitation was observed when thereconstituted sample was left at room temperature for 8 hours,indicating that the docetaxel albumin nanoparticle pharmaceuticalcomposition of the present invention was physically stable.

Example 3

An organic phase was obtained by adding 10 ml of ethanol into 150 mg ofdocetaxel, which was then dissolved by sonication. Recombinant humanserum albumin and bovine serum albumin solids (1:1 w/w) were dissolvedwith water for injection to formulate a 10 mg/ml solution of recombinanthuman serum albumin and bovine serum albumin 50 mg of glutathione wasadded to 450 ml of the 10 mg/ml solution of recombinant human serumalbumin and bovine serum albumin, and the solution was incubated in awater bath at 60° C. for 6 minutes to obtain an aqueous phase. Theorganic phase was homogeneously dispersed into the aqueous phase athigh-speed shear (1,000 revolutions per minute (rpm)), and the resultingsystem was transferred to an equipment for concentration byultrafiltration (PALL, 100 kD membrane). The system was concentrated byultrafiltration to obtain a 6 mg/ml concentrate of docetaxel. Prolinewas then added to the concentrate, such that the weight ratio of prolineto docetaxel was 1:1. The docetaxel albumin nanoparticles had an averageparticle diameter of 110 nm (Malvern Nano-ZS90). The concentrateobtained was sterilized by filtration through a 0.22 μm sterile filterhead, and freeze-dried for 48 hours in a lyophilizer. The lyophilizedsample was reconstituted with physiological saline, and no precipitationwas observed when the reconstituted sample was left at room temperaturefor 8 hours, indicating that the docetaxel albumin nanoparticlepharmaceutical composition of the present invention was physicallystable.

Example 4

The product was prepared according to the process of Example 1, exceptethanol was replaced by tert-butyl alcohol, glutathione was replaced byacetylcysteine, and arginine was replaced by lysine (the weight rationof lysine to docetaxel was 1, i.e., was 1:1). The particle diameter ofthe docetaxel albumin nanoparticles was 124 nm.

Example 5

The product was prepared according to the process of Example 1, exceptethanol was replaced by a mixture of acetone and tert-butyl alcohol (1:1v/v), glutathione was replaced by mercaptoethanol, and arginine wasreplaced by cysteine (the weight ration of cysteine to docetaxel was 1,i.e., was 1:1). The particle diameter of the docetaxel albuminnanoparticles was 132 nm.

Example 6

The product was prepared according to the process of Example 1, exceptglutathione was replaced by a mixture of dithiothreitol andmercaptoethanol (1:1 w/w), arginine was replaced by a mixture ofarginine and proline (the weight ration of arginine and proline was 1:1,and the weight ration of the mixture of amino acids to docetaxel was 1,i.e., was 1:1), and the concentration of docetaxel after concentrationwas 1 mg/ml. The particle diameter of the docetaxel albuminnanoparticles was 200 nm.

Example 7

The product was prepared according to the process of Example 1, exceptno glutathione was added, and arginine was replaced by glutamic acid(the weight ration of glutamic acid to docetaxel was 1:1). The particlediameter of the docetaxel albumin nanoparticles was 183 nm.

Example 8

The product was prepared according to the process of Example 2, exceptno glutathione was added, and proline was replaced by glutamic acid (theweight ratio of glutamic acid to docetaxel was 1:1). The particlediameter of the docetaxel albumin nanoparticles was 196 nm.

Comparative Example 1

An organic phase was obtained by adding 150 mg of docetaxel into a 50 mlvial, followed by addition of 10 ml of ethanol, and docetaxel wasdissolved by sonication. A concentrated solution (200 mg/ml) of humanserum albumin was diluted with water for injection to formulate a 6mg/ml solution. 50 mg of glutathione was added to 125 ml of the 6solution of human serum albumin, and the solution was incubated in awater bath at 80° C. for 6 minutes. The organic phase was thenhomogeneously dispersed into the aqueous phase at high-speed shear(1,000 revolutions per minute (rpm)), and the resulting system wastransferred to an equipment for concentration by ultrafiltration (PALL,100 kD membrane). The system was concentrated by ultrafiltration toobtain a 10 concentrate of docetaxel. Tartaric acid (salt) was thenadded, such that the final solution had a pH of 7.0, and containeddocetaxel in a concentration of 6 mg/ml, and tartaric acid (salt) in aconcentration of 100 mM (reference: the preferred ratios inCN103054798A). The sample of the final solution was sterilized byfiltration through a 0.22 μm sterile filter head, and freeze-dried for48 hours in a lyophilizer.

Comparative Example 2

The product was prepared according to the process of Comparative Example1, except tartaric acid was replaced by citric acid, and the finalconcentration was 100 mM (reference: the preferred ratios inCN103054798A).

Comparative Example 3

The product was prepared according to the process of Comparative Example1, except tartaric acid was replaced by sodium citrate, and the finalconcentration was 100 mM (reference: the preferred ratios inCN103054798A).

Comparative Example 4

The product was prepared according to the process of Example 1, exceptno amino acid was added.

Experimental Example 1: Storage Stability

After being left at 2-8° C. for various periods (see Table 1), theproducts of Examples 1 to 8 and Comparative Examples 1 to 4 were addedwith appropriate amounts of a 0.9% sodium chloride solution. After theproducts were uniformly dispersed, about 300 μl of each of the sampleswas accurately weighed, placed in a 2 ml centrifuge tube, and 600 μl ofacetonitrile was added accurately. After being vortexed for 30 seconds,the samples were subjected to solid phase extraction, and the resultingextraction solutions were filtered, so as to facilitate the detection ofthe content of 7-epi-docetaxel.

The contents in percentage of 7-epi-docetaxel were determined by HPLC,and the chromatographic conditions were as follows: column: SpherisorbRP18 4.6×250 nm, particle diameter: 5 μm; mobile phase: solutionA=acetonitrile:water (2:3, V/V), and solution B=acetonitrile; elutionwas carried out with solution A for the first 70 minutes, and then with10% solution A and 90% solution B for 20 minutes; flow rate: 1 ml/min;detection wavelength: 227 nm; and injection volume: 20 μl.

The increments (represented by peak area percentages in the HPLCchromatograms) of 7-epi-docetaxel in the docetaxel albumin nanoparticlepharmaceutical compositions after storage at 2-8° C. is shown in Table1.

TABLE 1 The increase of the content of 7-epi-docetaxel (increment % =the peak area % after N months − the peak area % on day 0) increment %of 7-epi-docetaxel Group 3 months 6 months 12 months 24 months 30 monthsComparative 0.45% 0.81% 1.54% 2.65% N/A Example 1 Comparative 0.44%0.86% 1.39% 2.56% N/A Example 2 Comparative 0.43% 0.85% 1.36% 2.47% N/AExample 3 Comparative 0.45% 0.87% 1.49% 2.79% N/A Example 4 Example 1-a0.02% 0.14% 0.37% 0.58% 0.74% Example 1-b 0.01% 0.11% 0.34% 0.49% 0.61%Example 1-c 0.04% 0.07% 0.30% 0.41% 0.54% Example 2 0.02% 0.15% 0.37%0.70% N/A Example 3 0.04% 0.16% 0.32% 0.63% N/A Example 4   0% 0.19%0.37% 0.66% N/A Example 5 0.07% 0.26% 0.43% 0.64% N/A Example 6 0.03%0.14% 0.39% 0.53% N/A Example 7 0.03%  0.2% 0.43% 0.63% N/A Example 8 0.7% 0.24% 0.46% 0.79% N/A Note 1: The acceptable limit for thestability of the docetaxel albumin nanoparticle composition (i.e., thehighest content of 7-epi-docetaxel) is normally “a content percentage of7-epi-docetaxel ≤1.0%”; Note 2: N/A means no detection was performed.

The experimental results showed that the addition of arginine, lysine,proline, cysteine, glutamic acid or a combination thereof to thedocetaxel albumin nanoparticle composition as prepared in the examplesof the present application can effectively inhibit the degradation ofdocetaxel, and reduce the increase of the content of the epimer,7-epi-docetaxel. It was also shown in Table 1 that the addition of anorganic acid (or a salt thereof) with pKa of 2.5 to 4.5 cannot inhibitthe increase of the content of the epimer, 7-epi-docetaxel.

In the above tests, the products of Examples 1-a, 1-b and 1-c of thepresent invention, wherein arginine was employed as an impurityinhibitor, achieved the best effects: it can be stored at 2-8° C. for atleast 24 months, or even up to 30 months.

Experimental Example 2: Storage Stability

Docetaxel albumin compositions comprising no arginine or comprisingarginine and docetaxel in a weight ratio of 80:1 were prepared, andcontent percentages of 7-epi-docetaxel were determined according toExperimental Example 1. The peak area percentages in the HPLCchromatograms over time are shown in Table 2. As can be seen from Table2, when the ratio of arginine to docetaxel was 80, the increase of thecontent of 7-epi-docetaxel in the composition was inhibited during 30months storage at 2-8° C.

TABLE 2 The increase of the content of 7-epi-docetaxel (increment % =the peak area % after N months − the peak area % on day 0) Ratio ofarginine to increment % of 7-epi-docetaxel docetaxel 3 months 6 months12 months 24 months 30 months 0 (0:1) 0.47% 0.87% 1.56% 2.76% 4.23% 80(80:1) 0.01% 0.04% 0.22% 0.38% 0.49%

As can be seen from Experimental Examples 1 and 2, the docetaxel albuminnanoparticles prepared in the examples of the present application havegood physical and chemical stability, are not liable to degradation,have less impurity after conventional storage and transportation, andcertainly would have less toxic and side effects caused by the impurity.

Experimental Example 3: Antitumor Activity

The use of the compositions prepared in the present application for thetreatment of cancer was demonstrated by a pharmacological efficacyverification test on a tumor model of BALB/c nude mice with asubcutaneous xenograft of human non-small cell lung cancer cell A549.

Thirty qualified BALB/c animals bearing a tumor of A549 cells wereselected, and randomly divided into three groups (10 mice per group),which were a group administered with physiological saline (blankcontrol), a group administered with a commercially available docetaxelinjection (20 mg/kg) and a group administered with the docetaxel albuminnanoparticle (20 mg/kg) according to Example 1-c of the presentinvention, respectively. The administration was performed throughintravenous injection via tail vein for 4 weeks. During theadministration, general clinical symptoms of the animals were observedtwice a day, the body weight and tumor diameter were measured twice aweek, and the tumor weight was measured at the end. The tumor size wasplotted versus time, so as to obtain a curve for the evaluation ofpharmacological efficacy.

The pharmacological efficacy results are shown in FIG. 1. As shown inFIG. 1, the docetaxel albumin nanoparticle pharmaceutical compositionaccording to Example 1-c of the present invention achieved a significantinhibitory effect on tumor growth on the tumor model of BALB/c nude micewith a subcutaneous xenograft of non-small cell lung cancer cell A549,and the effect of the docetaxel albumin nanoparticle compositionaccording to Example 1-c of the present invention was better that thatof the commercially available docetaxel injection.

Experimental Example 4: Antitumor Activity

The use of the compositions prepared in the present application for thetreatment of cancer was demonstrated by a pharmacological efficacyverification test with the docetaxel albumin nanoparticle pharmaceuticalcomposition of Example 7 of the present invention on a tumor model ofBALB/c nude mice with a subcutaneous xenograft of human non-small celllung cancer cell A549.

Thirty qualified BALB/c animals bearing a tumor of A549 cells wereselected, and randomly divided into three groups (10 mice per group),which were a group administered with physiological saline (blankcontrol), a group administered with a commercially available docetaxelinjection (16 mg/kg) and a group administered with the docetaxel albuminnanoparticle (13.6 mg/kg) according to Example 7 of the presentinvention, respectively. The administration was performed throughintravenous injection via tail vein for 4 weeks. During theadministration, general clinical symptoms of the animals were observedtwice a day, and the body weight and tumor diameter were measured twicea week.

The pharmacological efficacy results are shown in Table 3. The docetaxelalbumin nanoparticle pharmaceutical composition according to Example 7of the present invention achieved a significant inhibitory effect ontumor growth on the tumor model of BALB/c nude mice with a subcutaneousxenograft of non-small cell lung cancer cell A549. After administeredfor 21 days, the inhibition rate of tumor achieved by the docetaxelalbumin nanoparticle composition of Example 7 (administered dose: 13.6mg/kg) was 112%, while the inhibition rate of tumor achieved by thecommercially available docetaxel injection (administered dose: 16 mg/kg)was 99%. It can be seen that the docetaxel albumin nanoparticle of thepresent invention achieved a high inhibition rate of tumor at a lowdose, had a therapeutic effect significantly better than that of thecommercially available docetaxel injection, and had an excellenttherapeutic effect on human non-small cell lung cancer.

TABLE 3 The therapeutic effect on nude mice with a subcutaneousxenograft of human non-small cell lung cancer cell A549 Adminis- TumorTumor Inhibition tration volume volume rate of dose on day 0 on day 21tumor Group (mg/kg) (mm³) (mm³) (%) Physiological saline — 149.2 1218.9— Composition of Example 13.6 152.2 133.7 112 7 Commercially available16 141.2 151.5 99 docetaxel injection

Experimental Example 5: Antitumor Activity

The use of the compositions prepared in the present application for thetreatment of cancer was demonstrated by a pharmacological efficacyverification test with the docetaxel albumin nanoparticle pharmaceuticalcomposition of Example 7 of the present invention on a tumor model ofBALB/c nude mice with a subcutaneous xenograft of human ovarian cancercell SK-OV-3.

Fifty qualified BALB/c animals bearing a tumor of SK-OV-3 cells wereselected, and randomly divided into five groups (10 mice per group),which were a group administered with physiological saline (blankcontrol), group 1 administered with a commercially available docetaxelinjection (9.9 mg/kg), group 2 administered with a commerciallyavailable docetaxel injection (16 mg/kg), group 1 administered with thedocetaxel albumin nanoparticle (8.5 mg/kg) according to Example 7 of thepresent invention, and group 2 administered with the docetaxel albuminnanoparticle (13.6 mg/kg) according to Example 7 of the presentinvention, respectively. The administration was performed throughintravenous injection via tail vein for 4 weeks. During theadministration, general clinical symptoms of the animals were observedtwice a day, and the body weight and tumor diameter were measured twicea week.

The pharmacological efficacy results are shown in Table 4. The docetaxelalbumin nanoparticle pharmaceutical composition according to Example 7of the present invention achieved a significant inhibitory effect ontumor growth on the tumor model of BALB/c nude mice with a subcutaneousxenograft of ovarian cancer cell SK-OV-3. After administered for 18days, the inhibition rate of tumor achieved by the docetaxel albuminnanoparticle composition of Example 7 (administered dose: 8.5 mg/kg) was65%, while the inhibition rate of tumor achieved by the commerciallyavailable docetaxel injection (administered dose: 9.9 mg/kg) was 55%.When the pharmaceutical composition of Example 7 of the presentinvention was administered at a dose of 13.6 mg/kg and the commerciallyavailable docetaxel injection was administered at a dose of 16 mg/kg,the tumor inhibition rate were both 87% after administration for 18days. According to the above experimental results, compared with thecommercially available docetaxel injection, the composition of thepresent invention achieved a better or equivalent inhibitory effect ontumor at a lower dose, which demonstrated that the docetaxel albuminnanoparticle of the present invention had an excellent therapeuticeffect on human ovarian cancer.

TABLE 4 The therapeutic effect on nude mice with a subcutaneousxenograft of human ovarian cancer cell SK-OV-3 Adminis- Tumor TumorInhibition tration volume volume rate of dose on day 0 on day 18 tumorGroup (mg/kg) (mm³) (mm³) (%) Physiological saline — 146.7 1876.3 —Composition of Example 8.5 145.0 758.1 65 7 (group 1) Commerciallyavailable 9.9 144.0 930.3 55 docetaxel injection (group 1) Compositionof Example 13.6 145.3 361.8 87 7 (group 2) Commercially available 16145.8 373.8 87 docetaxel injection (group 2)

Although the present invention has been further described through theabove specific examples, it should be understood that it is not limitedthereby. The present invention encompasses general aspects of the abovedisclosures, and a person skilled in the art is able to make variousmodifications or change various details of the present invention withoutdeparting from the spirit and scope of the present invention. Thus, thepresent description is presented for purposes of illustration only andnot by way of limitation.

The invention claimed is:
 1. A storage stable docetaxel albuminnanoparticle pharmaceutical composition comprising: docetaxel, albumin,and amino acid(s), wherein the weight ratio of albumin to docetaxel isfrom 50:1 to 1:1; wherein the weight ratio of the amino acid(s) todocetaxel is from 1:1 to 20:1; wherein the amino acid(s) is selectedfrom the group consisting of arginine, lysine, proline, cysteine,glutamic acid and mixtures thereof; and wherein the amino acid(s)inhibits the formation of the 7-epi-docetaxel impurity of docetaxel inthe pharmaceutical composition thereby forming a storage stablepharmaceutical composition.
 2. The pharmaceutical composition accordingto claim 1, wherein the amino acid(s) is the only stabilizer in thepharmaceutical composition.
 3. The pharmaceutical composition accordingto claim 1, wherein the pharmaceutical composition optionally comprisesa protein structure unfolding agent, which includes one or more ofmercaptoethanol, glutathione, and acetylcysteine, and wherein the weightratio of albumin to the protein structure unfolding agent is from 50:1to >0:1.
 4. The pharmaceutical composition according to claim 1, whereinthe pharmaceutical composition is a nanoparticle suspension, whichcomprises docetaxel in a concentration of at least 1 mg/ml, and whereinthe nanoparticles therein have a particle diameter of not greater than200 nm.
 5. The pharmaceutical composition according to claim 1, whereinthe albumin includes one or more of recombinant albumin and serumalbumin.
 6. A method for preparing the pharmaceutical compositionaccording to claim 1, wherein the method comprises the followingsteps: 1) dissolving docetaxel in an organic solvent preferably selectedfrom one or more of ethanol, tert-butyl alcohol, and acetone, to obtainan organic phase; 2) using an aqueous solution containing amino acid(s)to dissolve albumin or to dilute a solution of albumin, so as to obtaina solution of albumin and amino acid(s); 3) subjecting the solution ofalbumin and amino acid(s) obtained in step 2) to an incubation reaction;4) adding the organic phase obtained in step 1) to the solution obtainedafter the incubation reaction in step 3) under shear, to obtain a dilutesolution of docetaxel albumin nanoparticles; and 5) concentrating thesolution obtained in step 4) by ultrafiltration, to obtain a docetaxelalbumin nanoparticle pharmaceutical composition with enhanced stability;wherein the enhanced stability is achieved by the amino acid(s) throughinhibiting the formation of the 7-epi-docetaxel impurity in thedocetaxel albumin nanoparticle pharmaceutical composition; wherein theweight ratio of albumin to docetaxel is not greater than 50, i.e., isfrom 50:1 to >0:1, preferably is from 50:1 to 1:1, and more preferablyis from 20:1 to 1:1; wherein the weight ratio of the amino acid(s) todocetaxel is no less than 0.5, i.e., is ≥0.5:1, preferably is from 0.5:1to 80:1, more preferably is no less than 1, i.e., is ≥1:1, andpreferably is from 1:1 to 20:1; and wherein the amino acid(s) preferablyincludes at least one of arginine, lysine, proline, cysteine, andglutamic acid, and preferably is arginine.
 7. The method according toclaim 6, wherein step 3) further comprises adding a protein structureunfolding agent to the solution of albumin and amino acid(s) obtained instep 2) before performing the incubation reaction; wherein the proteinstructure unfolding agent includes one or more of mercaptoethanol,glutathione, acetylcysteine, and dithiothreitol, and wherein the weightratio of albumin to the protein structure unfolding agent is not greaterthan 100:1, i.e., is from 100:1 to >0:1, and preferably is from 50:1to >0:1.
 8. The method according to claim 6, wherein alternatively,steps 2) and 3) are performed as follows: using water for injection todissolve albumin or to dilute a solution of albumin, so as to obtain asolution of albumin, and then adding amino acid(s) before subjecting thesolution to an incubation reaction.
 9. The method according to claim 6,wherein alternatively, steps 2) and 3) are performed as follows: usingwater for injection to dissolve albumin or to dilute a solution ofalbumin, so as to obtain a solution of albumin; adding amino acid(s) anda protein structure unfolding agent to the solution, and performing anincubation reaction; wherein the protein structure unfolding agentincludes one or more of mercaptoethanol, glutathione, acetylcysteine,and dithiothreitol; and wherein the weight ratio of albumin to theprotein structure unfolding agent is not greater than 100:1, i.e., isfrom 100:1 to >0:1, and preferably is from 50:1 to >0:1.
 10. The methodaccording to claim 6, wherein alternatively, steps 2) and 3) areperformed as follows: using water for injection to dissolve albumin orto dilute a solution of albumin, so as to obtain a solution of albumin,and subjecting the solution of albumin to an incubation reaction; andstep 5) is performed as follows: concentrating the solution byultrafiltration before adding amino acid(s) to the concentrate, toobtain a docetaxel albumin nanoparticle pharmaceutical composition withenhanced stability.
 11. The method according to claim 6, whereinalternatively, steps 2) and 3) are performed as follows: using water forinjection to dissolve albumin or to dilute a solution of albumin, so asto obtain a solution of albumin, adding a protein structure unfoldingagent to the solution, and performing an incubation reaction; andwherein step 5) is performed as follows: concentrating the solution byultrafiltration before adding amino acid(s) to the concentrate, toobtain a docetaxel albumin nanoparticle pharmaceutical composition withenhanced stability; wherein the protein structure unfolding agentincludes one or more of mercaptoethanol, glutathione, acetylcysteine,and dithiothreitol, and wherein the weight ratio of albumin to theprotein structure unfolding agent is not greater than 100:1, i.e., isfrom 100:1 to >0:1, and preferably is from 50:1 to >0:1.
 12. Aformulation comprising the docetaxel albumin nanoparticle pharmaceuticalcomposition according to claim 1, wherein the formulation furthercomprises a pharmaceutically acceptable carrier and/or auxiliarymaterial; and wherein the formulation is an injectable solution,injectable suspension, injectable emulsion, or lyophilized powderformulation.
 13. A formulation comprising the docetaxel albuminnanoparticle pharmaceutical composition prepared according to the methodof claim 10, wherein the formulation further comprises apharmaceutically acceptable carrier and/or auxiliary material; andwherein the formulation is an injectable solution, injectablesuspension, injectable emulsion, or lyophilized powder formulation, andpreferably is a lyophilized powder formulation.
 14. A method for thetreatment or prevention of an abnormal cell proliferative disease ordisorder, wherein the method comprises administering the pharmaceuticalcomposition according to claim 1 to a subject in need thereof.
 15. Amethod for the treatment or prevention of cancer, wherein the cancerincludes prostate cancer, gastric cancer, colon cancer, breast cancer,head and neck cancer, pancreatic cancer, lung cancer and ovarian cancer,wherein the method comprises administering the pharmaceuticalcomposition according to claim 1 to a subject in need thereof.
 16. Amethod for inhibiting the formation of the 7-epi-docetaxel impurity in adocetaxel albumin nanoparticle pharmaceutical composition, wherein themethod comprises adding amino acid(s) to the pharmaceutical composition;the amino acid(s) preferably includes at least one of arginine, lysine,proline, cysteine, and glutamic acid, and more preferably the aminoacid(s) is arginine.
 17. The method according to claim 16, wherein thepharmaceutical composition comprises: docetaxel, albumin, and aminoacid(s), wherein the weight ratio of albumin to docetaxel is not greaterthan 50, i.e., is from 50:1 to >0:1, preferably is from 50:1 to 1:1, andmore preferably is from 20:1 to 1:1; wherein the weight ratio of theamino acid(s) to docetaxel is no less than 0.5, i.e., is ≥0.5:1,preferably is from 0.5:1 to 80:1, more preferably is no less than 1,i.e., is ≥1:1, and preferably is from 1:1 to 20:1; and wherein the aminoacid(s) includes at least one of arginine, lysine, proline, cysteine,and glutamic acid, and preferably is arginine.
 18. A method forimproving the stability of a docetaxel albumin nanoparticlepharmaceutical composition, wherein the method comprises adding aminoacid(s) to the docetaxel albumin nanoparticle pharmaceuticalcomposition, so as to inhibit the formation of the 7-epi-docetaxelimpurity therein, thereby improving the stability; the amino acid(s)preferably includes at least one of arginine, lysine, proline, cysteine,and glutamic acid, and more preferably the amino acid(s) is arginine.19. The method according to claim 18, wherein the pharmaceuticalcomposition comprises: docetaxel, albumin, and amino acid(s), whereinthe weight ratio of albumin to docetaxel is not greater than 50, i.e.,is from 50:1 to >0:1, preferably is from 50:1 to 1:1, and morepreferably is from 20:1 to 1:1; wherein the weight ratio of the aminoacid(s) to docetaxel is no less than 0.5, i.e., is ≥0.5:1, preferably isfrom 0.5:1 to 80:1, more preferably is no less than 1, i.e., is ≥1:1,and preferably is from 1:1 to 20:1; and wherein the amino acid(s)includes at least one of arginine, lysine, proline, cysteine, andglutamic acid, and preferably is arginine.
 20. The pharmaceuticalcomposition according to claim 1, wherein the weight ratio of albumin todocetaxel is from 20:1 to 1:1.
 21. The pharmaceutical compositionaccording to claim 1, wherein the amino acid is arginine.
 22. Thepharmaceutical composition according to claim 1, wherein the albumin ishuman serum albumin.